초록
Sustainable production of biochemicals from cellulosic biomass is an attractive alternative to chemical production from fossil fuels. We describe the further development of a three protein synthetic pathway to convert NADPH and H<SUP>+</SUP> to hydrogen using a ferredoxin-NADP<SUP>+</SUP> reductase, a ferredoxin, and the [FeFe] hydrogenase from Clostridium pasteurianum at a rate greater than 14 mmol H<SUB>2</SUB> L<SUP>-1</SUP> hr<SUP>-1</SUP> using natural enzymes. We also demonstrate the feasibility of coupling this pathway to a cell-free extract to convert glucose to hydrogen in a potentially cost effective manner. Both of these accomplishments serve as the basis for further engineering to optimize both the yield and productivity of a low-cost cell-free process for the production of highly reduced biochemicals.